Mutants of non-essential genes are therefore readily obtained, facilitating identification of essential genes.
The in vitro and in vivo effects of licD2 mutation observed here correlate with those seen with licD2 mutants of non-typeable H. influenzae [4], [39].
Another important observation was that the deletion/substitution mutants of non-aromatic residues such as Gln2 and Pro3 did not show any comprehensive change in stability under all of the conditions tested.
Also, sapD mutants of non-typable H. influenzae have been shown to be attenuated in a chinchilla model of otitis media [87]. A. pleuropneumoniae possesses a complete sap operon, which could have significant role in the survival of the pathogen in the host.
Of the 1,711 queries, 1,377 are deletion mutants of non-essential genes and 334 are essential gene alleles.
The development of these methods should enable us to construct null mutants of non-model self-incompatible plants to examine the effects of the candidate genes on self-incompatibility signaling.
Of the 3985 mutants of non-essential genes in the collection thus examined, 110 showed a reduction in biofilm formation nine of which have not been well characterized yet.
To determine whether SAGA subunits outside the DUB module affected the cells' requirement for APC/C function, we tested whether deletion mutants of non-DUB SAGA components could suppress APC/C mutants.
We generated a set of mutants of non-expanded RUNX2 with enhanced CC propensity by inserting canonical CC-stabilizing hydrophobic residues (V and L) in positions a/d of two heptads within the CC heptad register established by the valine residues that immediately precede and follow the polyQ/polyA region (Fig. 3A; Supplementary Material, Fig. S1E).
We have found that polyA-expanded RUNX2 has some toxicity when overexpressed in cells, similar to what was observed with other polyA-expanded proteins (3), and that cc+ mutants of non-expanded RUNX2 are as toxic, or even more toxic, than polyA-expanded RUNX2.
A cc− mutant of non-expanded, wt RUNX2 was also generated [i.e. RUNX2 cc−); Fig. 3A and B].
